Targeted transgenic mice using CRISPR/Cas9 Technology

Abstract

CRISPR/Cas9 is a powerful technology that has transformed gene editing of mammalian genomes, being faster and more cost-effective than standard gene targeting techniques. In this chapter, we provide a step-by-step protocol to obtain Knock-Out (KO) or Knock-In (KI) mouse models using CRISPR/Cas9 technology. Detailed instructions for the design of single guide RNAs (sgRNA) for KO approaches and single-strand oligonucleotide (ssODN) matrix for generation of KI animals are included. We also describe two independent CRISPR/Cas9 delivery methods to produce gene-edited animals starting from zygote-stage embryos, based either on cytoplasmic injection or electroporation.